• Korean Journal of Plant Resources
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• v.21 no.2
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• pp.134-138
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• 2008

Nerine from south africa and its sparkling flower shape make us estimate it as a hopeful kind of cut follow. There was a few studies on Nerine in korea. We started this study to set bulb propagation methods. The propagation by tissue culture was changeable according to the growth regulators The best growth regulator combination which makes a lot of Bulblet was NAA $0{\sim}0.5$ + BA $0.5{\sim}2.0mg\;{\cdot}\;L^{-1}$ in Nerine bowdenii ‘Favourite’ and Nerine sarniens ‘Red’ respectively. The adjust culture media source for tissue culture were glucose 9% as a carbon source and ($NH_4+NO_3$) 40mM as a nitrogen source. When glucose was used as a carbon source, Bulblet were harvested a little bit low then sucrose but comparative emergence rate was so high that it is good for carbon source in nerine tissue culture. When we consist culture media as MS+BA $1.0mg\;{\cdot}\;L^{-1}$+sucose 7% + ($NH_4+NO_3$) 40mM, the produced Bulblet were reached up to 1.7 each per bulb and emergence rate was up to 100% irrespective of acclimatization period. The suitable culture explant for nerine tissue culture was scale. When scale was cultured with MS+BA $1.0mg\;{\cdot}\;L^{-1}$+sucose 7%, its propagation efficiency was 54 times greater than using growing point. A proper culture part of the scaly leaf was middle part (8 scaly leaf from outer 8th scaly leaf) when middle part was cultured the number of Bulblet were up to 1.8 each per explant.

• KSBB Journal
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• v.19 no.6 s.89
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• pp.471-477
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• 2004

Oldenlandia diffusa is a Chinese medicinal herb with antitumor activity capable of suppressing the growth of some cancer cell lines. Oleanolic acid and ursolic acid are triterpenoid compounds that exist in Oldenlandia diffusa. Recently, these have been noted for anti-inflammatory, anti-cancer, and hepato-protective effects. Application of both plant growth regulators, 2,4-D and kinetin, was found to be essential for the initiation of callus and suspension cells. Leaf blades of Oldenlandia diffusa was transformed into callus on Schenk and Hildebrandt medium supplemented with 0.5 mg/L 2,4-D and 0.1 mg/L kinetin, while optimum initiation condition for suspension cells of Oldenlandia diffusa was determined to be 0.75 mg/L 2,4-D and 0.1 mg/L kinetin. Chromatographic separation of oleanolic acid from its derivatives was achieved using Rexchrom S5-100-ODS column. Analytical conditions for oleanolic acid were determined as follows: flow rate at 1.0 mL/min, UV length at 200 nm and mobile phase of $80\%$ acetonitrile and $20\%$ water. Production of secondary metabolites was found to be increased by the treatment with elicitors or signal transducers. The maximum production of oleanolic acid was 99.6 mg/L in cultures with 0.5 mM salicylic acid. It is 1.74 times higher than that of control.

• Journal of Korean Society of Forest Science
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• v.13 no.1
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• pp.25-39
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• 1971

Recently successful induction of haploid plant by means of anther culture method has become a big topic among geneticists and plant breeders. The haploid plant can be used as a precious material for such basic researches as mutation or genetics. Once the haploid is obtained, production of homozygous plant is not a difficult problem. The method of producing homozygous plant can, also, be applied to the practical breeding works. When applied to the hybridization of self-fertilizing breeding period would be greatly shortened and in cross-fertilizing vegetables production of uniform hybrid seed would be very easily obtained. Last few years many scientists attempted anther cultures using various plant species, but it was successful only in several species. Unlike the other tissue cultures which use somatic organs or tissues as explants, anther culture seems to be very difficult because the plants or calli have to be induced from the haploid microspores or pollen grains. In the present experiment anther culture of fruit trees and ornamental shrubs of four genera and seven species was attemped. Anthers of Various stages ranging from tetrad and late microspore were cultured on the modified Murashige and Skoog's medium supplemented with various concentrations of auxins and kinetin as growth regulators. Handling of materials, sterilization, and other operations of culture were done by routine methods. The results were summarized as follows: 1. Calli were induced in the anthers of Forsythia Koreana Nak., Rhododendron mucronuratum Turcz., R. yedoense Max. var. Poukhanense Nak., and Prunus armeniaca L. var. ansu Max. No signs of callus were observed in Prunus persica Sieb. et Zucc. var. vurgaris Max., Pyrus ussuriensis var. macrostipes (Nak.), and Prunus salcina Lindley. 2. Calli were easily formed in any of the media with differing concentrations of auxins and kinetin. 3. In F. Koreana calli developed from anther surface and connective. Callus emerging out of anther locule was not observed. 4. Somatic calli arose from filament, connective, and inside of anther wall in R. mucronulatum. Many of the microspores accumulated starch grains. 5. The anther lobes located opposite the filament of R. yedoense turned easily to calli. This phenomenon was not observed in R. mucronulatum. Microspore embedded for a period in the medium became starch pollen. No callus was observed arising from microspore. 6. In P. armeniaca calli were not induced from somatic anther tissues. Instead, callus emerged out of anther locule rupturing the anther slit. Starch was not formed in the microspore. 7. In P. persica, Pyrus ussuriensis, and P. salcina, calli were not observed in the anthers examined more than 60 days after culture. Microspores of these species, however, were free of starch grains even after long period of subculture. 8. It was learned that somatic calli of the species examined arose usually from endothelium of anther wall, septum of two neighboring anther locules, parenchyma tissues of connectives, or anther lobes. 9. In the anther locule of P. armeniaca cultured long in medium, swollen microspores, polynucleate microspores, multicellular pollen grains, or callus mass were frequently observed, this indicating that the callus of this species was microspore-origin. 10. It was clarified that in P. armeniaca production of haploid plant by anther culture might be possible.

• Journal of Plant Biotechnology
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• v.41 no.3
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• pp.116-122
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• 2014

Total of fifty accessions of Brassica rapa with various morphological characteristics were used for production of double haploid plants though microspore culture in Brassica rapa. Among them, only 30 accessions induced embryos from microspores. The highest efficiency of embryo induction of 1.194 per bud was obtained from IT135449 of turnip type, while 3 accessions of sarson (winter oil) type did not generate embryo. The effect of heat shock periods for embryogenesis was also investigated with 4 accessions (IT135449; Turnip type, IT199710; Chinese cabbage type, IT212886; Pak choi type, IT218043; Summer oil type). The high productions of embryos were observed in IT135449, IT199710 and IT212886 when microspores were pre-cultured to $32^{\circ}C$ for 2 days. In IT218043, high embryogenesis was observed at the 3 days of heat shock treatment. The optimal condition of shoot regeneration for IT199710 was observed in MS medium supplemented with NAA $0.5mg{\cdot}L^{-1}$ and BAP $1mg{\cdot}L^{-1}$. In contrast, the IT135449 and IT212886 were observed high regeneration frequency in MS medium without plant growth regulators. All the plantlets regenerated from microspore-derived embryos have been successfully transplanted to soil, and bud self-pollinated seeds were produced from doubled haploid plants. This indicated that double-haploid genotype was likely generated naturally during embryogenesis process.

• Journal of Korean Society of Forest Science
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• v.43 no.1
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• pp.6-13
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• 1979

In order to substract the time and cost of propagation for inducing the haploid plants per each species. 500 anthers of late uninucleate microspore on early binucleate microspore stage of Robinia pseudoacacia (Fuel tree) Punius granatum (Ornamental tree). Aleurites fordii (Faty tree) and Styrax japonica (Silvicultural tree) were cultured on the modified Murashige and Skoog's medium supplemented with Kinetin, 2.4-D and NAA as growth regulators. And I observed the samples of cultured anthers under the microscope which were made by Microtoming method and Paraffin method. The results were summarized as follows: 1) Among 500 cultured anthers per each species, anther numbers inducing the diploid callus were as follow: Styrax japonica 20 (4% for the species total); Aleurites fordii 10 (2% for the species, total) and Punica granatum 45 (9% for the species total) were showed. 2) 2n Callus were induced from anther wall. but haploid callus were induced from anther locule. 3) Haploid callus were induced only in 25 anthers (5% for the species total) of Robinia pseudoacacia. 4) These haploid callus were not originated from body cell of anther wall tissue, but from reduced microspores, 5) Since already reported many herbaceous haploid plants were induced from the callus which were originated from reduced microspores, I conclude that the anther of woody plant which induced the haploid callus also will be cultured haploid plant.

• Korean Journal of Medicinal Crop Science
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• v.1 no.2
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• pp.137-157
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• 1993

This study was carried out to investigate the optimal condition for multiple propagation through leaf tissue culture and to apply anther culture techniques to Pulsatilla koreana Nakai breeding. Leaf and anther of Pulsatilla koreana Nakai were cultured on MS, MT, LS and $B_5$ media supplemented with several growth regulators and nitrogen sources under various conditions. For callus induction and differentiation from the Pulsatilla koreana leaf segments were more effective in the combination of zeatin and auxin than auxin alone. The color of the callus was green when treated with IBA alone. Shoot differentiation was more effective when treated with zeatin than auxin alone, especially the best hormoal combination for shoot differentiation was zeatin 1.0mg/l +NAA 0.1mg/l, while 2,4-D inhibited shoot differentiation. The appeared rate of S pollen was 35% in vivo, while that of S pollen by low temperature$(4^{\circ}C)$ pretreatment for 4 days was increased by 53% and the optimum culture time for callus induction from anther was uni-nucleate stage. $B_5$ basal medium supplemented with NAA 0.5mg/l and zeatin 1 mg/l was the most effective on callus formation and the best results of plant regeneration were obtained from combination of NAA 0.5mg/l and zeatin 0.5mg/l in anther culture. $NH_{4}NO_3$ as more effectives as the nitrogen source than $KNO_3$ and the combination with zeatin 2.0mg /L was the best effective. The best combination for plant regeneration in callus induced from anther was $NH_{4}NO_3$ 1650mg/l + $KNO_3$ 3800mg/l + zeatin 2.0mg/l. Ploidy level of anther-derived plants appeared 28% haploid, 47% diploid and the others were triploid, tetraploid and mixploid. In compare with E.S.T, M.D.H and P.X banding patterns were distinguished among callus, haploid and diploid plants in electrophoresis.

• Journal of Bio-Environment Control
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• v.23 no.2
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• pp.77-82
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• 2014

The present study was conducted to establish an effect and a proper concentration for treatment with gibberellic acid ($GA_3$) and thidiazuron (TDZ), resulting with increase berry size and yield in Gaeryangmeoru grapes. Berry size was increased by treatment with $GA_3$, and the fruit clusters obtained for the groups treated with $GA_3$ concentrations of 100 and $200mg{\cdot}L^{-1}$ were bigger. The berry number was also enhanced in $GA_3$ treated groups, but the soluble solid content and acidity was not significantly different. Damage caused by $GA_3$ treatment, such as peel pollination and berry shatter, was observed in the group with $200mg{\cdot}L^{-1}$. The berry size was larger in group treated with a high concentration of $GA_3$ and TDZ respectively than in those treated with low concentrations in the treatment mixed $GA_3$ and TDZ; however, fruit with low soluble solid content and high acidity was harvested after $GA_3$ and TDZ treatment due to delay of berry ripening. The pericarp tissue layers were not changed, but the distance from the epidermis layer to vascular bundle tissue was increased as a result of $GA_3$ and TDZ treatment. Therefore, $GA_3$ and TDZ did not affect an cell division but not cell size, resulting in an enlarged berry size. It is necessary to treat plant growth regulators 2~3 times and immediately after berry set to enhance berry set rate, because the period of berry set is short. This study suggests that the proper concentration for enhancing berry size and set were up to $100mg{\cdot}L^1$ $GA_3$ or $50mg{\cdot}L^{-1}GA_3+1.25mg{\cdot}L^{-1}$ TDZ, and it is necessary to pay attention to harvest mature fruits because of the delay of ripening caused by the usage of TDZ.

• KOREAN JOURNAL OF CROP SCIENCE
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• v.40 no.6
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• pp.757-767
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• 1995

This experiment was done to determine the optimum concentration of IAA for root development in plants regenerated from the callus culture of barley embryos. Two concentrations of 2,4-D, 3ppm and 5ppm selected as an optimum among five different concentrations in the previous experiment were used for callus induction and proliferation in this experiment. For callus induction, 3ppm of 2,4-D produced 35.6% in immature embryos and 4.4% in mature embryos, while 5ppm gave 33.8% in immature and 5.6% in mature embryos. Out of 320 immature embryos cultured, 111 embryos were induced to calli and 684 plants were produced from them, while only 16 embryos were induced to calli from 320 mature embryos and 92 plants were restored. The rates of callusing and plant regeneration were 34.7%, 214% in immature embryos and 5.0%, 28.7% in mature embryos, respectively. The average root lengths and root numbers of plants restored from callus at five different IAA concentrations of 0ppm, Ippm, 5ppm, l0ppm and 30ppm were 7.9mm, 3.6; 18.4mm, 5.2; 16.1mm, 3.9; 8.5mm, 3.5 and 6.4mm, 3.4, while plants directly obtained from mature embryos were 14.8mm, 4.9; 4.9mm, 3.6; 4.3mm, 3.1; 3.6mm, 2.6 and 3.2mm, 2.1, respectively. Therefore, 1ppm gave the best result for the root. promotion in callus, whle 0ppm, a control, gave the largest root developmemt in embryos. High concentration of lAA(30ppm) in callus and any exogeneous supplement of lAA in embryos negatively affected to the root lengths and root numbers. Genotypic effect was also observed in given four varieties, Bruce, Klages, Olbori and Albori. For chromosome doubling, when 0.1% colchicine was applied on 428 plants under three different conditions such as air circulation, temperatures and growth stages, 319 plants of doubled haploids were obtained so that the rate was 74.5%

• KOREAN JOURNAL OF CROP SCIENCE
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• v.51 no.spc1
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• pp.160-165
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• 2006

Biofuctionality of soybean seeds and soy-bean products have been fortified by the uncovering of the multifuctional beneficial effects of isoflavones. As one way to fully utilize beneficial effects of isoflavones in soybean sprout is through the enhancement of isoflavone contents in soybean seeds, genetic selection for higher isoflavone and cultivational measures to increase isoflavone content in soybean seeds were attempted. Isoflavones (daidzein, gemstein) contents in soybean seeds and soybean sprouts were determined by high performance liquid chromatography. Total isoflavone contents in soybean seeds ranged from 756 to $1,682{\mu}g/g$ and Iksan #13 $(1,682{\mu}g/g)$ showed highest content among the 21 germplasms analyzed. Onetime treatment of soybean plants with Antipol or Piaster at the $V_4$ stage yielded seeds with higher isoflavones as $2,472{\mu}g/g\;or\;2,052{\mu}g/g$, respectively, which were higher by 37% and 14% than that of seeds in the control plants, respectively. In Eunhakong, Isoflavone contents of soybean sprout changed during sprouting. Daidzein content in hypocotyl increased to maximum on the third day of cultivation and decreased there-after, whereas the content changed little in cotyledon. In sprouts of Pungsannamulkong, daidzein content in hypocotyl showed a maximum level on the first day and decreased gradually thereafter but, the content changed little in cotyledon. Total isoflavone contents in lateral roots which developed on the 6th day after sprouting ranged from 4,416 to $5,232{\mu}g/g$ DW.

• Korean Journal of Agricultural Science
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• v.9 no.2
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• pp.467-483
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• 1982

Leaflets of Allium sativum L. (garlic) and leaf scales Allium cepa L. (onion) were cultured on the Murashige and Skoog(MS) medium to study the effects of plant growth regulators, temperature and light on the formation of shoot and root. And also, the potentiality of shoot or root formation of tissues in accordance with the leaf age and the region was discussed. In the experiment with garlic, the formation of shoot and root was more effective on the medium containing $BA\;2mg/{\ell}$ and $NAA\;1mg/{\ell}$ and it occured on the basal region of outer leaflets. The shoot formation was more effective at $20^{\circ}C$ than $28^{\circ}C$ under 16-hour daylength, and it was more effective in darkness than light condition at $28^{\circ}C$. The results of shoot and root formation on the new cloves of garlic harvested in 1982 were similar to those the old cloves of garlic harvested in 1981. In the experiment with leaf seales of onion, when the tissues of dormant leaf scales of onions harvested in 1981 were explanted on the MS medium, a few shoots were formed on the medium containing $BA\;2mg/{\ell}$ and $NAA\;1mg/{\ell}$. In this case,there were no differences on the shoot formation among the age and region of leaflets. On the culture of leaf scales, the shoot was not formed on the leaf scales located at outer or middle region, but it was formed on the medium containing BA accompanied with NAA at inner scales. On the medium complemented with BA alone, the tissues were not differentiated, but on the medium complemented with NAA, the callus was formed from tissues and then the root was formed through the callus.